DJ-1 interacting proteins analyzed by tandem affinity purification mass spectrometry / 中国医师杂志

Objective:RNA interference technology (siRNA) was used to inhibit the expression of DJ-1 gene in lung squamous cell carcinoma HTB-182 cells, then, tandem affinity purification mass spectrometry (TAP-MS) was performed to screen the interacting proteins of DJ-1 in lung cancer cell line of HTB-182.Methods:The siRNA lentivirus vector targeting DJ-1 gene was constructed to infect HTB-182 cells (DJ-1 siRNA group), and the lentivirus vector control group (control siRNA group) and blank control group were established. The expression level of DJ-1 protein was detected by Western blot, and the endogenous DJ-1 protein silenced si-DJ-1-HTB-182 cells were established. The specific primers of DJ-1 were designed, and the DJ-1 expression plasmid pNTAP-DJ-1 with streptomycin binding peptide label (SBP) and calmodulin binding peptide label (CBP) was constructed. The cell line DJ-1 siRNA HTB-182 was stably transfected with liposome, and the positive clones were screened by G418. The positive clones were verified by Western blot, and the interacting proteins of DJ-1 were found by TAP-MS.Results:The protein expression of DJ-1 in DJ-1 siRNA interference group was significantly lower than that in empty plasmid group and blank control group ( P<0.05); HTB182 cell line stably expressing pNTAP-DJ-1 plasmid was successfully constructed; Three proteins interacting with DJ-1 were screened by TAP-MS: cytokeratin 1 (keratin 1), cytokeratin 10 (keratin 10) and NADPH oxidase activating protein P47 (P47 Px). Conclusions:Keratin 1, Keratin l0 and P47 Px protein may be DJ-1 interactions protein.

Clinical Comparation of Two Kinds of Chemotherapy Regimens in the Treatment of Patients with MDS-RAEB/AML-MRC / 中国实验血液学杂志

OBJECTIVE@#To investigate the influence of conventional CAG regimen and decitabine + decreased dose CAG (D+dCAG) regimen on the clinical efficacy and safety of patients with MDS-RAEB/AML-MRC.@*METHODS@#The clinical data of 67 patients with MDS-RAEB/AML-MRC hospitalized in our hospital from March 2012 to July 2017 were analyzed retrospectively. According to chemotherapecctic regimens, 76 patients were divided into 2 groups: 37 patients treated with conventional CAG regimen were enrolled in control group, 30 patients treated with decitabine + decreased dose CAG regimen were enrolled in D+dCAG group. The complete remission (CR) rate, overall remission rate (ORR), OS and PFS time and incidence of adverse reactions in 2 groups were compared.@*RESULTS@#The CR in D+dCAG group was significantly higher than that in control group (P＜0.05). ORR was not significanly different between 2 groups (P＞0.05). There was no significant difference in the cumulative OS rate between 2 groups (P＞0.05). There was no significant difference in the cumulative OS rate and PFS rate in nonimplantation between 2 groups (P＞0.05). The incidence of adverse reactions of hematological system, pulmonary infection, skin and soft tissue infection, agranulocytosic fever and mycotic infection was not significanly different between 2 groups (P＞0.05). The duration of granulocyte deficiency and platelet count less than 20×10/L were not significanly different between 2 groups (P＞0.05).@*CONCLUSION@#Compared with conventional CAG regimen, decitabine + decreased dose CAG regimen in the treatment of patients with MDS-RAEB/AML-MRC can efficiently improve the remission effects and showed the well overall safety, but can not increase the survival rate.

Correlation of Early Minimal Residual Disease Level with Prognosis in AML Patients with NPM1 Mutation Positive after Chemotherapy / 中国实验血液学杂志

OBJECTIVE@#To analyze the correlation of the minimal residual disease level with the prognosis of the AML patients with NPM1 gene mutation positive after chemotherapy.@*METHODS@#The clinical data of 112 newly diagnosed adult AML patients with positive NPM1 gene were collected and retrospectively analyzed. The correlation of the transcripts of NPM1 gene mutation with prognosis of patients was analyzed.@*RESULTS@#In 112 AML patients, the median transcript level of NPM1 gene mutation accounted for 83.68% (5.86%-486.57%), FLT3-ITD mutation positive was found in 44 cases (39.29%), chromosomal abnormalities in 22 cases (19.64%) and complete remission in 96 cases (85.71%). Multivariate logistic regression analysis showed that the initial induction therapy and white blood cell count closely related with complete remission (P＜0.05). The median follow-up time was 22 (3-36) months, and the 3-year overall survival rate was 66.07% in 112 patients. Multivariate logistic regression analysis showed that both the high level of minimal residual disease at the initial complete remission and the high level of minimal residual disease after consolidation therapy were the independent risk factors for overall survival (P＜0.05).@*CONCLUSION@#In newly diagnosed adult AML patients with NPM1 mutation positive, the early high level of minimal residual disease after chemotherapy closely relates with poor prognosis.

Regulatory Effect of MiR-543 on Proliferation and Apoptosis of Human Leukemia Cell Line K562 and Its Mechanism / 中国实验血液学杂志

OBJECTIVE@#To investigate the regulatory effect of miR-543 on proliferation and apoptosis of human leukemia cell line K562 and its mechanism.@*METHODS@#46 CML patients treated in our hospital from 2018.2-2018.9 was selected and enrolled in CML group, and 30 healthy persons were selected and enrolled in control group at the same time. After Lipofectamine 2000 liposome was used to transfer the miR-543 mimic and negative control (Scramble mimic) to K562 cells, CCK8 assay was used to detect the effect of miR-543 on proliferation of K562 cells; Luciferase reporter assay was used to report the targeting relationship of miR-543 to Wnt gene. Flow cytometry was used to detect the effect of miR-543 on apoptosis of K562 cells; Western blot method was used to detect the Wnt, β-catenin, BCL-2, c-MYC and BAX expression level.@*RESULTS@#The level of miRNA-543 in CML patients was significantly lower than that in healthy controls (P＜0.05). The expression level of miR-543 in mimic group was significantly higher than that in blank control group (P＜0.05). The Wnt gene expression level in mimic group was significantly lower than that in blank control group (P＜0.05). The expression of luciferase of Wnt wild plasmid was decreased by miR-543 mimic (P＜0.05), and the luciferase activity of Wnt mutant plasmid was not inhibited by miR-543 mimic after mutation of binding site (P＜0.05). There was no significant difference in the proliferation ability of K562 cells between the blank control group and the negative control (Scramble mimic) group (P＞0.05). The proliferation level of K562 cells in mimic group was significantly lower than that in blank control group and negative control (Scramble mimic) group (P＜0.05). Apoptotic level of K562 cells in miR-543 mimic group was significantly higher than that in blank control group and negative control (Scramble mimic) group (P＜0.05). Western blot assay showed that Wnt, β-catenin, BCL-2 and c-MYC protein levels in miR-543 mimic group were significantly lower than those in blank control group and negative control (Scramble mimic) group (P＜0.05); BAX protein level in miR-543 mimic group was higher than that in blank control group and negative control (Scramble mimic) group (P＜0.05).@*CONCLUSION@#miR-543 can target Wnt protein to inhibit the activity of Wnt signaling pathway, thereby inhibiting the proliferation of leukemia cells and increasing the level of apoptosis.

A study on genetic polymorphisms of rs600231 and rs4102217 loci in promoter of MALAT1 gene in Guangxi population / 临床检验杂志

Objective@#To investigate the single nucleotide polymorphisms (SNPs) of rs600231A/G and rs4102217 G/C in the promoter region of MALAT1 (metastasis associated in lung adenocarcinoma transcript 1) gene in the healthy population of Guangxi district and analyze the differences in the population among different regions. @*Methods@#The genotypes of rs600231A/G and rs4102217G/C of 207 healthy individuals in Guangxi were detected by SNPscan high-throughput technique. The genotype and allele frequency distributions were analyzed statistically with the data of HapMap-CEU (European population), HapMap-HCB (Beijing Han population), HapMap-JPT (Japanese population) and HapMap-YRI (African population) published by Human genome Haplotype Map (HapMap). @*Results@#There were three genotypes of AA (38.2%), AG (46.4%) and GG (15.4%) in rs600231A/G, and the differences were significantly different compared with the polymorphism of Japan and Africa population (HapMap-JPT and HapMap-YRI) (P＜0.05). Compared with HapMap-CEU, the genotype difference was not statistically significant (P＞0.05), but the allele distribution was statistically different (P＜0.05). The rs4102217 G/C polymorphism contained GG(75.4%), CG(23.2%) and CC(1.4%), and the polymorphisms were significantly different from those in European and Japanese populations (P＜0.05). There was no significant difference between gender in the polymorphisms of the two loci (P＞0.05). @*Conclusion@#The polymorphisms of rs600231A/G and rs4102217G/C in the MALAT1 promoter region were found in Guangxi healthy population, and the distribution of polymorphisms may be different in the population of various regions. @*@#

Ratio of Primary Bone Marrow Cells after Induction Chemothrapy for Two Weeks in the Patients with PhALL and Its Influence on Complete Remission and Overall Progrosis / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the influence of bone marrow blasts ratio after induction chemotherapy for 2 weeks in patients with PhALL, and it's influence on complete remission (CR) and overall prognosis.</p><p><b>METHODS</b>A total of 172 patients with PhALL in our hospital from March 2012 to February 2016 were selected. The bone marrow blast ratio was analyzed by the receiver-operating characteristic curve (ROC) in patients after induction chemotherapy for 2 weeks, at same time its influence on CR and overall prognosis of PhALL patients was evaluated.</p><p><b>RESULTS</b>The cutoff value of CR was 0.075, its area under ROC was 0.763; the comparison of area under ROC with A=0.5 showed statistically significant difference, therefore 172 patients with PhALL were grouped according to bone marrow blast ratio after induction chemotherapy for 2 weeks: 104 cases (60.5%) with bone marrow blast ratio <0.075, 68 cases (39.5%) with bone marrow blast ratio ≥0.075. The PhALL patinets with bone marrow blast ratio <0.075 who achieved CR and finally achieved CR after induction chemotherapy for 4 weeks acconnted for 89 (85.6%) and 99(95.2%) respectively, which were significantly higher than those in PhALL patients with bone marrow blast ratio≥0.075, [29(42.6%) and 52 (76.5%)](P<0.05). In addition, the influencing factor clinically reducing the OS and DFS rate of patients and enhancing the ralapse rate of patients were mainly chemotherapy, the failure of induction chemotherapy (patients did not achieve CR after induction therapy for 4 weeks), the bone marrow blast ratio≥0.075 after induction treatment for 2 weeks, and CNSL at diagnosis and so on, while the enhaced WBC count at diagnosis was poor factor affecting the DFS rate of patients.</p><p><b>CONCLUSION</b>After induction chemotherapy for 2 weeks, the elevated bone marrow blast ratio in PhALL patients will be infavourable to CR, and the overall prognosis is poor.</p>

Study of genetic polymorphisms of miR-146a,miR-149 gene in Guangxi popula-tion / 中国免疫学杂志

Objective:To study the frequencies of allele and genotype distribution of miR-146a C>G(rs2910164) and miR-149 T>C( rs2292832) gene, and to analyze the statistical differences between different racial and nationalities.Methods:The Polymerase Chain Reaction-Single Base Extension ( PCR-SBE) technique and DNA sequencing methods were used for the determination of the SNP in miR-146a C>G and miR-149 T>C gene,and compared with the European, African, Japanese and People in Beijing from the Human Genome Project (HapMap).Results:There were no statistical differences of allele and genotype distribution in miR-146a C>G,miR-149 T>C between female and male group (P>0.05).There were significant difference frequencies of allele and genotype distribution of miR-146a C>G and miR-149 T>C gene by compared with the European, African and People in Beijing( PG and miR-149 T>C in Guangxi populations, and there were significant differences by compared with other ethnic populations, which may play an important role in the human inherited disease research.

Risk factors of erectile dysfunction in patients with cardiovascular diseases / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the penile erectile function of hospitalized male patients with cardiovascular diseases, the incidence of erectile dysfunction (ED) in this cohort, and the relationship of ED with cardiovascular diseases and its risk factors.</p><p><b>METHODS</b>Using a self-designed questionnaire, we conducted an investigation among the hospitalized patients in the Department of Cardiovascular Diseases of the First and Second Affiliated Hospitals of Xi'an Jiaotong University. We measured their body height, body mass index (BMI), waist circumference, hip circumference, and blood pressure, obtained their personal data, past history, metabolic indexes, and erectile function scores by IIEF-5, and analyzed the risk factors of ED using univariate and multivariate logistic regression and OR analyses.</p><p><b>RESULTS</b>Totally, 225 valid questionnaires were included in this investigation, which showed a 66.7% incidence of ED, 15.8% mild, 27.0% mild to moderate, 17.6% moderate, and 6.3% severe. The incident rates of ED in the 18-35 yr, 36-49 yr, 50-65 yr, and > 65 yr age groups were 13.6%, 39.1%, 89.2%, and 91.2%, respectively. Univariate logistic regression analysis manifested that the risk factors of ED in the patients with cardiovascular diseases included age (OR = 3.122, 95% CI 2.040-4.779), smoking (OR = 1.768, 95% CI 1.209-2.584), BMI (OR = 1.261, 95% CI 1.114-1.427), total cholesterol (OR = 1.77, 95% CI 1.339-2.340), TC/HDL (OR =1.715, 95% CI 1.349-2.181), hypertension (OR = 1.717, 95% CI 1.110-2.658), and coronary heart disease (OR = 2.235, 95% CI 1.169-4.275), while multivariate logistic regression analysis showed the risk factors to be age (OR = 4.99, 95% CI 2.264-10.998), financial condition, (OR = 2.804, 95% CI 1.127-6.976), smoking (OR = 2.109, 95% CI 1.179-3.772), BMI (OR = 1.414, 95% CI 1.136-1.760), and TC/HDL (OR = 2.001, 95% CI 1.016-3.943).</p><p><b>CONCLUSION</b>The incidence of ED is high in hospitalized patients with cardiovascular diseases and rises with the increase of age. Age, smoking, financial condition, BMI, and TC/HDL are the risk factors of both ED and cardiovascular diseases, and financial condition is closely associated with ED.</p>

miR-139-5p and inhibits invasion and metastasis of hepatoma cells by targeting TGF-β1 / 中华肝胆外科杂志

Objective To investigate the molecular mechanism of miR-139-5p targeting transforming growth factor-β1 (TGF-β1) in the regulation of epithelial mesenchymal transition (EMT),thus inhibiting invasion and metastasis of hepatoma cells.Methods Bioinformatics methods were used to determine whether miR-139-5p was the best binding miRNA of TGF-β1.Correlation between the TGF-β1 expression as detected by immunohistochemistry and Western blot,and the miR-139-5p level by qRT-PCR in 56 hepatoma tissues and 20 normal tissues,respectively,was analyzed.The relationship between the miR-139-5p level as detected by qRT-PCR,and TGF-β1,E-cadherin and Vimentin by Western blot in the high and low metastatic hepatoma cell lines were investigated.In recombinant cell lines,whether miR-139-5p could bind to the 3'UTR site of TGF-β1 was evaluated,and the effect on invasive ability after modulating miR-139-5p level was also tested by the transwell method.Results A total of 20 miRNAs were found to be able to bind with TGF-β1 by bioinformatics methods and among these mRNAs,miR-139-5p was the best target miRNA with the highest specificity and strongest stability to bind TGF-β1.The positive expression rates of TGF-β1 in hepatoma tissues and adjacent normal liver tissues were 80.4％ (45/56) and 15.0％ (3/20),respectively,(P ＜0.05).There were significant differences on the expressions of TGF-β1,E-cadherin and Vimentin among the different metastatic cell lines (all P ＜ 0.05).After miR-139-5p was transfected into hepatoma cells,miR-139-5p could bind to the 3'UTR site of TGF-β1,resulting in downregulating TGF-β1 expression.When compared to the other three groups,the cell line with a high expression of miR-139-5p had a significantly lower count of invasive cells (53 ± 4/high magnification field) (P ＜ 0.05).Conclusion miRNA139-5p could specifically bind to the 3'UTR site of TGF-β1 and regulate the EMT signaling pathway,thus suppressing invasion and metastasis of hepatoma cells.

A novel schiff base zinc coordination compound inhibits proliferation and induces apoptosis of human osteosarcoma cells / 华中科技大学学报（医学）（英德文版）

Various kinds of schiff base metal complexes have been proven to induce apoptosis of tumor cells. However, it remains largely unknown whether schiff base zinc complexes induce apoptosis in human cancer cells. Here, we synthesized a novel schiff base zinc coordination compound (SBZCC) and investigated its effects on the growth, proliferation and apoptosis of human osteosarcoma MG-63 cells. A novel SBZCC was synthesized by chemical processes and used to treat MG-63 cells. The cell viability was determined by CCK-8 assay. The cell cycle progression, mitochondrial membrane potential and apoptotic cells were analyzed by flow cytometry. The apoptosis-related proteins levels were determined by immunoblotting. Treatment of MG-63 cells with SBZCC resulted in inhibition of cell proliferation and cell cycle arrest at G1 phase. Moreover, SBZCC significantly reduced the mitochondrial membrane potential and induced apoptosis, accompanied with increased Bax/Bcl-2 and FlasL/Fas expression as well as caspase-3/8/9 cleavage. Our results demonstrated that the synthesized novel SBZCC could inhibit the proliferation and induce apoptosis of MG-63 cells via activating both the mitochondrial and cell death receptor apoptosis pathways, suggesting that SBZCC is a promising agent for the development as anticancer drugs.

Study of the relationship among expression of Survivin and MRP and the drug resistance in human nasopharyngeal carcinoma / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#This study aimed to explore the relationship among expression of Survivin and MRP and drug resistance in NPC.@*METHOD@#Expression of Survivin were detected by immunohistochemistry method in 45 cases of NPC and 24 cases of normal mucous membrane of nasopharynx (NMMN). The relationship between expression of Survivin and pathological factors in NPC were analysized. Expression of Survivin and MRP were detected in 31 patients of NPC with paclitaxel resistance and 20 patients of NPC without paclitaxel resistance. The relation- ship among the expression of Survivin or MRP and paclitaxel resistance in NPC were analysized. The paclitaxel resistance cell line, 5-8F-PTX(+); was established by a step-increased method. The expression of Survivin and MRP were detected by western blot in 5-8F-PTX(+) and 5-8F.@*RESULT@#The positive were 71. 1% (32/45) in NPC and 8.33% (2/24) in NMMN. And there were significantly differences between them (P < .05). There were relationship among expression of Survivin and differentiation degree, lymph node metastasis, distant metastasis, and clinic stages of NPC. The positive were 75.9% (31/39) in moderately differentiated NPC and 16.7% (1/6) in lowly differentiated NPC, respectively. There were significantly differences between them (P < 0.05). The positive of Survivin were 83.9% (26/31) in NPC patients with paclitaxel resistance and 45.0% (9/20) in NPC patients without Paclitaxel resistance, respectively. There were significantly differences between them (P < 0.05). The positive of MRP were 87.1% (27/31) in NPC patients with paclitaxel resistance and 40.0% (8/20) in NPC patients without paclitaxel resistance, respectively. There were significantly differences between them (P < 0.05). There were positive correlation between the expression of Survivin and MRP in NPC patients with Paclitaxel resistance. The expression of Survivin and MRP were higher in 5-8F-PTX(+) than in 5-8F. The IC50 of paclitaxel, cDDP, 5-FU and Vincristine were significantly higher in 5-8F-PTX(+) than in 5-8F.@*CONCLUSION@#There were relationship among the expression of Survivin and difference, metastasis and TNM stages of NPC. Survivin may serves as a molecular marker for development and progress in NPC. There were relationship among the high expression of Survivin and MRP and increasing of drug resistance in NPC.

Research on chemotherapy measures in different Clonorchis sinensis infectios-ity endemic areas / 中国血吸虫病防治杂志

Objective To explore the suitable chemotherapy measures in different Clonorchis sinensis infectiosity endemic ar-eas. Methods The interventional groups and control groups were set up in the heavy,moderate and light C. sinensis infectiosity areas in Guangdong Province,respectively. In the intervention groups,the chemotherapy was administered among all the residents aged above 3 years in the heavy endemic areas,the chemotherapy was given among the focus populations in the moderate endemic areas,and the chemotherapy was administered among the residents infected with C. sinensis confirmed by stool examinations in the light endemic areas. No measures were carried out in the control groups. Results One year after the interventions,the C. sinensis infection rates of the interventional groups decreased by 47.90%,86.52%and 100%,the abnormal rates of liver B ultrasonic ex-aminations decreased by 26.50%,31.00%and 100%,the ALT abnormal rates decreased by 48.70%,62.30%and 100%,and the AST abnormal rates decreased by 55.50%,39.90% and 100% in the heavy,moderate and light endemic areas,respectively. There was no decreasing trend of above-mentioned indexes in the control groups. Conclusion In the different C. sinensis infectios-ity endemic areas,the different chemotherapy measures should be carried out.

Efficacy of spinal pia mater incision and laminoplasty combined with internal fixation for old spinal cord injury / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To evaluate the clinical efficacy of incising spinal pia mater to relieve pressure and unilateral open-door laminoplasty with internal screw fixation for treatment of the dated spinal cord injury.</p><p><b>METHODS</b>From March, 2009 to July, 2010, 16 cases with chronic cervical cord injury underwent spinal dura mater incision and unilateral open-door laminoplasty with internal screw fixation. Nerve functions of pre- and postoperation were evaluated by Frankel classification and the Japanese Orthopaedic Association (JOA) scale.The improvement rate of JOA score at the indicated time was recorded.</p><p><b>RESULTS</b>Postoperative Frankel classification rating of 16 patients improved obviously.JOA scores at the 1st month, 3rd month, 6th month, and 12th month after surgery were 7.9 ± 2.3, 8.5 ± 1.6, 8.9 ± 2.1, and 12.4 ± 2.5, respectively, and significantly increased compared with that prior to surgery (5.5 ± 0.6). At the end of follow-up period, JOA score was significantly higher than that of pre-treatment (P<0.05). The recovery was relatively rapid during the first 3 months following the surgery, then entered a platform period.</p><p><b>CONCLUSION</b>It is effective for patients with dated spinal cord injury to undergo spinal decompression and laminoplasty.</p>

Clinical safety about repairing the peripheral nerve defects with chemically extracted acellular nerve allograft / 中华外科杂志

<p><b>OBJECTIVE</b>To discuss the clinical safety about repairing the peripheral nerve defects with the acellular allogeneic nerve.</p><p><b>METHODS</b>The 41 patients (male 38, female 3, age 10 - 55 years old, average 28.9 years old) who were performed chemically extracted acellular nerve allograft transplanting to repair nerve defects from 2002 to 2011. The average interval from injury to nerve repairing was 4.1 months (range, 10 hours to 9 months). There were 41 cases nerve defects including 10 brachial plexus nerves, 3 radial nerves of upper arm, 4 ulnar nerves of forearm, 12 digital and toe nerves, 2 sciatic nerves, 2 femoral nerves, 3 tibial nerves and 5 common peroneal nerves. There were 12 cases combined fractures and 20 soft tissue injury or defects. The average length of the nerve allograft to bridge the nerve defects was 6.1 cm (range, 2 - 10 cm). No immunosuppressive drugs were used in all cases. The clinical safety was evaluated through physical examination, blood biochemistry and immunity detection.</p><p><b>RESULTS</b>All cases were followed up post-operation. They got primary wound healing except 2 superficial infection who got delay healing through dressings changing. No any adverse effects happened including immunological rejection, hypersensitivity reaction, deep infection, hepatotoxicity and nephrotoxicity.</p><p><b>CONCLUSIONS</b>It is safe and feasible to repairing human peripheral nerve defects with chemically extracted acellular nerve allograft.</p>

Screening for methylation-silenced genes in acute myeloid leukemia HL-60 cell line by a quantitative proteomic approach / 中南大学学报(医学版)

Objective To screen for new methylation association genes in HL-60 to reveal the pathogenesis of leukemia, and provide important theoretical and scientific basis for the prevention and cure of leukemia. Methods Two-dimensional fluorescence difference gel electrophoresis (F-2D-DIGE) was performed to separate the total proteins from acute myelogenous leukemia (AML) cell line HL-60 cells with or without 5-aza-2-deoxycytidine (5-aza-2-dC) treatment. Imaging software Decyder 6.5 and PDQuest were used to detect the differential expression protein spots, and matrix-assisted laser desorption/ionizaion time-of-flight mas spectrometer (MALDI-TOF MS) was adopted to identify the differential expression proteins. Results F-2D-DIGE maps of 5-aza-2-dC-untreated HL-60 and-treated HL-60 cells were established. A total of 53 differential protein spots were detected, and 35 differential proteins were successfully identified. Of the identified proteins, 32 proteins were up-regulated, and 3 proteins were down-regulated in HL-60 cells after 5-aza-2-dC treatment.Conclusion Thirty-five differential proteins may be associated with methylation in HL-60 cell line, which provides the important clues for epigenetic study of leukemia.

Methylation and expression of glioma pathogenesis-related protein 1 gene in acute myeloid leukemia / 中南大学学报(医学版)

OBJECTIVE@#To detect the methylation and expression of glioma pathogenesis-related protein 1(GLIPR1) gene in the acute myeloid leukemia (AML) cell lines and bone marrow cells from AML patients, and to determine the relationship between promoter methylation and expression of GLIPR1.@*METHODS@#Five leukemia cell lines, 54 bone marrows from the newly diagnosed AML patients, 48 bone marrows from the acute lymphoblastic leukemia (ALL )patients, 40 bone marrows from the chronic myeloid leukemia (CML) patients,35 bone marrows from control patients, and 8 bone marrows from the complete remission AML patients were collected. RT-PCR and methylation-PCR (MSP) were used to detect the mRNA expression and promoter methylation of GLIPR1, respectively, and the relationship between them was analyzed.@*RESULTS@#The level of GLIPR1 mRNA in the AML cell lines was lower than that in the chronic myeloid leukemia (CML) and ALL cell lines, whereas the methylation level of GLIPR1 in the former was higher than that in the latter. The level of GLIPR1 mRNA in the AML cell lines was significantly increased, but had no obvious changes in the CML and ALL cell lines after 5-aza-2dC treatment. The mRNA level of GLIPR1 in the AML bone marrows (0.38+/-0.20)was obviously lower than that in the ALL bone marrows (0.76+/-0.18), CML bone marrows (0.80+/-0.14), and control bone marrows(0.85+/-0.12). The level of GLIPR1 mRNA in the bone marrows with complete remission AML was obviously higher than that in the AML bone marrows before the treatment (0.78+/-0.13 vs. 0.36+/-0.20); but there was no obvious difference between the ALL bone marrows and the control bone marrows, and the CML bone marrows and the control bone marrows (both P>0.05). The positive rate of GLIPR1 gene methylation in the AML bone marrows (81.5%) was obviously higher than that in the ALL bone marrows(37.5%), CML bone marrows (27.5%) and the control bone marrows(14.3%). The positive rate of GLIPR1 gene in the bone marrows with complete remission AML was obviously lower than that in the bone marrows before the treatment (12.5% vs. 75.0%), but there was no obvious difference between the ALL bone marrows and between the control bone marrows,and the CML bone marrows and the control bone marrows (both P>0.05). There was a negative correlation between the mRNA level and methylation status of GLIPR1 in the AML bone marrows.@*CONCLUSION@#GLIPR1 expression is downregulated or even lost by promoter methylation in AML, and the expression and methylation level of GLIPR1 gene may have some significance in evaluating the curative effect of AML patients.

Analysis of in vitro anti-leukemia effect of 5-aza-2'-deoxycitydine / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effect of methylation transferase inhibitor 5-aza-2'-deoxycitydine (5-aza-2 dC) on the growth, differentiation and apoptosis of human acute myeloid leukemia(AML) cell line HL-60, and to explore the possible anti-leukemia mechanism of 5-aza-2 dC.@*METHODS@#HL-60 cells were treated by 5-aza-2 dC at various concentrations for different periods of time. The effect of 5-aza-2 dC on the growth of HL-60 cells were detected by MTT assay. The effect on the cell cycle and differentiation were detected by flow cytometry. The effect on the apoptosis were detected by Hochest33342 staining and flow cytometry. The expression of S100A8 and S100A9 was detected by reverse transcription-polymerase chain reaction (RT-PCR).@*RESULTS@#(1) 5-aza-2 dC inhibited the growth of HL-60 cells in a concentration- and time-dependent manner, and HL-60 cells were arrested at G2/M phases; (2) 5-aza-2 dC enhanced the expression of cell differentiation antigen CD11b at HL-60 cells, especially at the low drug concentration; (3) 5-aza-2 dC induced HL-60 cell apoptosis in a concentration- and time-dependent manner, especially at the high drug concentration; (4) 5-aza-2 dC increased the expression levels of S100A8 and S100A9 mRNA in HL-60 cells.@*CONCLUSION@#5-aza-2 dC can inhibit the growth of HL-60 cells accompanied with G2/M phase arrest, induce the differentiation and apoptosis of the cells, and increase the expression levels of S100A8 and S100A9 mRNA, which may be the anti-AML mechanism of 5-aza-2 dC.